Direct determination of free state low molecular weight compounds in serum by online TurboFlow SPE HPLC-MS/MS and its application
Direct determination of free state low molecular weight compounds in serum by online TurboFlow SPE HPLC-MS/MS and its application
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在线TurboFlow SPE HPLC-MS/MS直接测定血清中游离态低分子化合物及其应用
DOI：
10.1016/j.talanta.2018.10.082
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发表时间：
2018
期刊：
影响因子：
6.1
通讯作者：
Jiang Guibin
中科院分区：
文献类型：
--
作者：
Gao Ke;Fu Jianjie;Xue Qiao;Fu Jie;Fu Kehan;Zhang Aiqian;Jiang Guibin
血清中低分子量化合物（LMWCs）的游离浓度在环境健康和药代动力学领域都起着重要的作用。建立了在线TurboFlow SPE高效液相色谱-质谱联用技术直接测定血清中游离低分子肝癌的新方法。通过TurboFlow SPE柱去除大分子，包括蛋白质和蛋白质结合的lmwc。同时，在TurboFlow固相色谱柱上保留游离的LMWCs，用流动相洗脱，进行HPLC-MS/MS检测。选取与血清蛋白具有低、中、高亲和力的16种具有代表性的LMWCs进行方法优化和验证，其血清游离浓度检测结果与平衡透析法（r = 0.9714,p< 0.01）和超滤法（r = 0.9054,p< 0.01）检测结果基本一致。然后，将该方法成功地应用于人血清中游离lmwc的检测以及LMWC-HSA复合物的解离常数（Kd）的评估。本文首次对9Cl-PF3ONS、PFEtCHxS、8:2 diPAP等新兴污染物的kd值进行了评价。该方法处理时间快（18 min）、所需样品量小（25 μL）、操作方便（直接注射），可用于研究低分子肝细胞在血清中的生物利用度、生物蓄积消除、临床药物研发中用于治疗药物监测的游离低分子肝细胞的实时检测、血清中游离低分子肝细胞的降解动力学过程等。
The free concentration of low molecular weight compounds (LMWCs) in serum play important roles in both the environmental health and pharmacokinetic fields. A novel and broad spectrum method for direct determination of free LMWCs in serum was established by online TurboFlow SPE HPLC-MS/MS. Macromolecules, including proteins and protein-bound LMWCs, were removed by the TurboFlow SPE column. Meanwhile, free LMWCs were retained on the TurboFlow SPE column and eluted by the mobile phase for HPLC-MS/MS detection. 16 representative LMWCs with low, medium and high affinities to serum proteins were selected for method optimization and validation, and their free concentrations in serum detected by present method showed fairly consistent results with the equilibrium dialysis (r = 0.9714,p< 0.01) and ultrafiltration (r = 0.9054,p< 0.01) method. Then, present method was successfully applied to detect free LMWCs in human serum as well as to evaluate the dissociation constants (Kd) of LMWC–HSA complexes. Kdvalues of some emerging pollutants, such as 9Cl-PF3ONS, PFEtCHxS and 8:2 diPAP, were firstly evaluated by the present method. The rapid processing time (18 min), small sample amount requirement (25 μL) and convenient operation (direct injection) made present method be promising in studies on the bioavailability, bioaccumulation and elimination of LMWCs in serum, real-time detection of free LMWCs for therapeutic drug monitoring during clinical drug research and development, and the degradation kinetic processes of free LMWCs in serum.